[Construction of Escherichia coli gene knock-out mutants for engineering of fatty acid metabolism].

Autor:	Qu J; Institute of Microbiology Chinese Academy of Sciences, Beijing 100190, China. qujingqiu1986@163.com, Liu C, Liu W, Tao Y, Zhu K
Jazyk:	čínština
Zdroj:	Wei sheng wu xue bao = Acta microbiologica Sinica [Wei Sheng Wu Xue Bao] 2013 Jun 04; Vol. 53 (6), pp. 608-14.
Abstrakt:	Objective: Gene knock out technique is very important for gene function study. We developed a simple and efficient method to knock out chromosomal genes in Escherichia coli. Methods: Using the Escherichia coli Keio single-mutant library, we combined Red homology recombination with P1 phage transduction and developed a method to knock out the genes in Escherichia coli MG1655. Results: We obtained beta-oxidation mutants deltafadD, deltafadE and deltafadD-deltafadE and fatty acid synthesis mutants deltafabH, deltafabF and deltafabH-deltafabF. There were no obvious growth changes between deltafadD or deltafadE mutant strains and MG1655. However, deltafabH and deltafabH-deltafabF mutant strains grew much slower than the wild type strain. The fatty acid contents in deltafadD, deltafadE and deltafadD-deltafadE were 18.2 mg/L, 20.0 mg/L and 19.2 mg/L respectively, higher than 17.5 mg/L in wild type. The fatty acid contents in deltafabH, deltafabF and deltafabH-deltafabF were 12.6 mg/L, 15.2 mg/L and 11.2mg/L respectively, lower than that in wild type. Conclusion: Using Keio mutant library, P1 phage transduction and resistant gene elimination, we have established a simple and efficient method for gene knock-out in Escherichia coli.
Databáze:	MEDLINE
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